LOCUS       sequence-340588-        8550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutive expression of V5-tagged YAP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8550)
  AUTHORS   Ma X, Mandausch FJ, Wu Y, Sahoo VK, Ma W, Leoni G, Hostiuc M, 
            Wintgens JP, Qiu J, Kannaiyan N, Rossner MJ, Wehr MC
  TITLE     Comprehensive split TEV based protein-protein interaction screening 
            reveals TAOK2 as a key modulator of Hippo signalling to limit 
            growth.
  JOURNAL   Cell Signal. 2024 Jan;113:110917. doi: 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7.
  PUBMED    37813295
REFERENCE   2  (bases 1 to 8550)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Signal."; date: "2024-01"; volume: "113:110917. doi"; pages: " 
            10.1016/j.cellsig.2023.110917. Epub 2023 Oct 7"
COMMENT     Sequence Label: pLenti6_YAP1-V5
FEATURES             Location/Qualifiers
     source          1..8550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        1816..2119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2120..2323
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2274..2294
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2320..2344
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2440..2464
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3983..4007)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4060..4101
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4138..4158)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4143..4472
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4323..4458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4520..4567
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4586..4984
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5071..5304
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5376..5497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5413..5432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5467..5486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5537..5672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5692..5711)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5693..5711)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5721..5738)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5721..5737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5730..5752)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5879..6334
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5966..5985)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6176..6197
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6360..6464
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6465..6533,6534..7325)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6683..6702)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7496..8084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7985..8004
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8238..8255
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8372..8393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8408..8413,8414..8431,8432..8438)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8446..8462
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8451..8473
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8470..8486
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8470..8486
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8505..8525
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8507..8525
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgata
     1801 agcttgggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1861 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1921 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1981 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2041 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2101 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2161 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2221 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2281 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     2341 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gactctagag
     2401 gatccactag tccagtgtgg tggaattctg cagatatcaa caagtttgta caaaaaagca
     2461 ggctccacca tggatcccgg gcagcagccg ccgcctcaac cggcccccca gggccaaggg
     2521 cagccgcctt cgcagccccc gcaggggcag ggcccgccgt ccggacccgg gcaaccggca
     2581 cccgcggcga cccaggcggc gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc
     2641 cgcggggact cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg
     2701 gccaacgtgc cccagaccgt gcccatgagg ctccggaagc tgcccgactc cttcttcaag
     2761 ccgccggagc ccaaatccca ctcccgacag gccagtactg atgcaggcac tgcaggagcc
     2821 ctgactccac agcatgttcg agctcattcc tctccagctt ctctgcagtt gggagctgtt
     2881 tctcctggga cactgacccc cactggagta gtctctggcc cagcagctac acccacagct
     2941 cagcatcttc gacagtcttc ttttgagata cctgatgatg tacctctgcc agcaggttgg
     3001 gagatggcaa agacatcttc tggtcagaga tacttcttaa atcacatcga tcagacaaca
     3061 acatggcagg accccaggaa ggccatgctg tcccagatga acgtcacagc ccccaccagt
     3121 ccaccagtgc agcagaatat gatgaactcg gcttcaggtc ctcttcctga tggatgggaa
     3181 caagccatga ctcaggatgg agaaatttac tatataaacc ataagaacaa gaccacctct
     3241 tggctagacc caaggcttga ccctcgtttt gccatgaacc agagaatcag tcagagtgct
     3301 ccagtgaaac agccaccacc cctggctccc cagagcccac agggaggcgt catgggtggc
     3361 agcaactcca accagcagca acagatgcga ctgcagcaac tgcagatgga gaaggagagg
     3421 ctgcggctga aacagcaaga actgcttcgg caggcaatgc ggaatatcaa tcccagcaca
     3481 gcaaattctc caaaatgtca ggagttagcc ctgcgtagcc agttaccaac actggagcag
     3541 gatggtggga ctcaaaatcc agtgtcttct cccgggatgt ctcaggaatt gagaacaatg
     3601 acgaccaata gctcagatcc tttccttaac agtggcacct atcactctcg agatgagagt
     3661 acagacagtg gactaagcat gagcagctac agtgtccctc gaaccccaga tgacttcctg
     3721 aacagtgtgg atgagatgga tacaggtgat actatcaacc aaagcaccct gccctcacag
     3781 cagaaccgtt tcccagacta ccttgaagcc attcctggga caaatgtgga ccttggaaca
     3841 ctggaaggag atggaatgaa catagaagga gaggagctga tgccaagtct gcaggaagct
     3901 ttgagttctg acatccttaa tgacatggag tctgttttgg ctgccaccaa gctagataaa
     3961 gaaagctttc ttacatggtt agacccagct ttcttgtaca aagtggttga tatccagcac
     4021 agtggcggcc gctcgagtct agagggcccg cggttcgaag gtaagcctat ccctaaccct
     4081 ctcctcggtc tcgattctac gcgtaccggt tagtaatgag tttggaatta attctgtgga
     4141 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4201 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4261 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4321 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4381 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4441 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4501 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     4561 atacgacaag gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct
     4621 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     4681 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     4741 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     4801 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     4861 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     4921 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     4981 ctaagcacaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5041 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     5101 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5161 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5221 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     5281 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     5341 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     5401 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5461 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     5521 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5581 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5641 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     5701 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5761 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5821 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     5881 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     5941 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6001 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     6061 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     6121 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     6181 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     6241 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     6301 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     6361 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6421 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6481 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6541 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6601 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6661 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6721 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6781 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6841 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6901 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6961 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7021 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7081 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7141 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7201 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7261 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7321 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8161 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     8221 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     8281 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     8341 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8401 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8461 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
     8521 aaagggaaca aaagctggag ctgcaagctt
//
